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ISSN 0864-4551 (Print)

2018, Number 1

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Biotecnol Apl 2018; 35 (1)

Advantages of an immunomodulatory assay for the determination of the in vitro potency of human gamma interferon

Silega CLG, Gerónimo PH, Brito LDC, Bustamante PY, Delgado MI, Bello RI, Furrazola GG, Cruz GO, García IG, Izquierdo LM, Costa ALB

Language: English
References: 27
Page: 1301-1308
PDF size: 397.77 Kb.


ABSTRACT

Products containing human interferon human recombinant gamma interferon (IFNγ-hr) as active pharmaceutical ingredients are promising alternatives for treating juvenile rheumatoid arthritis (as Heberon® Gamma R) or skin cancer (e.g. HeberFERON®). Significantly, its development relies on the availability of analytical methods to evaluate the biological activity of the formulation. Therefore, an in vitro immunomodulatory assay was established for quantifying the rhIFNγ biological activity in the referred products, so as to fulfill the European Pharmacopoeia requirements. The average biological potency obtained for several batches using this assay was compared and against an antiviral assay in use. The antiviral assay is based on the rhIFNγ capacity of protecting Hep-2C cells against the viral attack. The immunomodulatory assay assesses the overexpression of Human Leukocyte Antigen-DR (HLA-DR) in COLO-205 cells when they are in contact to the IFNγ-hr. The absorbance readings were processed using a statistical parallel lines program. The immunomodulatory assay was specific for the rhIFNγ effect, with dose-response curves in a sensibility range 800-6.25 IU/mL, equivalent to the range 40-0.31 ng/mL. It was demonstrated that the recombinant human alpha-2b interferon (rhIFNα2b), present in the HeberFERON® formulation, does not interfere in the quantitative determination of rhIFNγ biological activity. Even though both assays detected similar values of biological activity for the different lots tested, the immunomodulatory assay results confirmed its advantages as compared to the antiviral assay.


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